二色胡枝子种子储藏蛋白多样性研究

采用SD-PAGE方法对二色胡枝子14个居群147个个体单粒种子盐溶蛋白进行了电泳检测,共检测到28个蛋白单体,其中多态性单体23个,多态百分率为82．14％,各居群内多态百分率平均为33％。各居群间的遗传分化系数（Gst）为0．4669。居群间基因流（Nm）为0．5709。种群内的基因多样性占总群体的53．3％,表明二色胡枝子种群具有丰富的遗传多样性,居群内变异是二色胡枝子遗传多样性的主要来源。聚类分析结果将14个二色胡枝子居群分为3类,居群间地理距离和遗传距离相关性不显著。     

浙江省桐庐县长叶榧居群遗传结构的ISSR分析

采用ISSR分子标记技术,对浙江省桐庐县白云源森林公园的6个长叶榧（Torreya jackii）亚居群进行分析,阐明长叶榧在小地理范围内的遗传结构。12个引物对6个长叶榧亚居群的120个个体进行扩增,共得到194个位点,其中多态位点87个,总多态位点百分率（P）为44.85%,平均为29.21%。长叶榧总Shannon信息指数（I）为0.166 3,平均为0.119 9;总Nei指数（h）为0.103 5,平均为0.075 5。P、I、h均表明长叶榧总体水平的遗传多样性较高,亚居群水平的遗传多样性较低。AMOVA分子变异分析显示,21.45%的变异存在于亚居群间,78.55%的变异存在于亚居群内。长叶榧亚居群间的遗传分化系数（G_st）为0.270 5,基因流为1.347 8。6个长叶榧亚居群的平均遗传距离为0.037 0,根据亚居群间的遗传距离进行UPGMA聚类,结果为生境相似的亚居群聚在一起。     

黄芩种质资源ISSR遗传多样性的分析及评价

采用ISSR分子标记技术对6个野生或栽培居群共147份黄芩种质进行遗传多样性分析和评价。分析结果表明,51个ISSR引物中筛选出18条扩增条带清晰、重复性好和多态性高的引物,共扩增出485条清晰的条带,其中466条具有多态性,平均多态性位点比率为96.08%,平均Nei’s基因多样性指数和Shannon’s信息指数分别为0.244 4和0.388 9,等位基因数（Na）和有效等位基因数（Ne）分别为1.993 8和1.383 9,遗传分化指数G_st=0.122 3,遗传一致度（I）和遗传距离（D）分别为0.951 5和0.050 1,说明收集的黄芩种质资源在总体上具有较高的遗传多样性,不同居群间存在一定的遗传分化和基因交流,遗传变异主要存在于居群内。分子聚类结果表明,同一地区的种质并没有按照收集来源完全聚类,可能与种质不同起源或民间栽培引种有关。在DNA分子水平揭示黄芩种质资源的遗传多样性水平,将为进一步黄芩种质资源评价、保存和新品种选育等利用提供依据。     

白花泡桐种源的遗传多样性和遗传分化研究

利用ISSR技术对白花泡桐38个种源的遗传多样性和遗传分化进行分析。结果显示：（1）从100个ISSR引物中筛选出9个能扩增出清晰带型并具多态性的引物,共扩增出95个条带,其中88条具多态性,多态位点比率为92.63%。（2）在物种水平上,有效等位基因数（N_e）、Nei’s基因多样性指数（H）、Shannon’s信息指数（I）的平均值分别为1.391 0、0.242 4、0.376 5;种源的多态位点比率在32.63%（江西抚州）~56.84%（广西梧州和江西九江）之间,平均为47.16%;基因流（N_m）为0.912 7,种源间遗传分化系数（G_st）为0.353 9,反映出种源间遗传变异占总遗传变异的35.39%,且遗传变异主要来源于种源内的个体间。（3）遗传一致度在0.39~0.82之间,反映出白花泡桐的遗传基础较宽。UPGMA法聚类分析将38个种源分为3组,主坐标分析(PCoA)将其大致分为4组,两种聚类方法的结果有一定的差异,本文做了相关讨论。研究还表明种源间的遗传距离与地理距离相关不显著。     

木荷种群在演替系列群落中的遗传多样性

利用ISSR分子标记对木荷种群在3个演替系列群落中的遗传多样性进行了研究。12个随机引物共检测到203个可重复的位点,其中多态位点183个,总多态位点百分率（P）为90.15%,平均多态位点百分率为82.27%。Shannon信息指数（I）估算的总遗传多样性为0.524 4,平均为0.477 8。Nei指数（h）计算的总基因多样性为0.358 7,平均为0.326 5。3个种群的P、I、h大小顺序均为针叶林>针阔混交林>常绿阔叶林。AMOVA分子变异显示91.56%变异来源于种群内,8.44%变异来源于种群间。种群间的遗传分化系数（G_ST）为0.089 7,基因流（N_m）为5.073 1。种群间的遗传相似度平均为0.928 4,遗传距离平均为0.074 4,针叶林种群与针阔混交林种群遗传相似度最高。     

二色胡枝子遗传多样性ISSR分析

利用ISSR分子标记技术对二色胡枝子的遗传多样性进行分析,16个ISSR引物共扩增出229条带,多态性条带209条,多态性比率为91%,居群的平均多态性位点比率为68.24%,胡枝子总遗传多样性Ht为0.392,Shannon指数为0.576,群体间基因分化系数Gst的变动范围非常大,平均值为0.2434,居群间基因流Nm为1.052。种群内的基因多样性占总群体的75.66%,种群间占24.34%,表明二色胡枝子种群具有丰富的遗传多样性,在育种上具有很大的遗传潜力。根据居群间遗传相似系数聚类,14个居群被聚成3大类群,且居群的遗传多样性参数与其地理、生态因子相关均不显著,遗传多样性无明显的地域性分布格局。     

滇牡丹遗传多样性的ISSR分析

应用ISSR标记对中国西南地区特有植物滇牡丹(Paeonia delavayi)的遗传多样性进行了研究。从100个引物中筛选出10个用于正式扩增,在取自16个自然居群和1个迁地保护居群的511个个体中,检测到92个多态位点。在居群水平上,多态位点百分率（PPB）为44.61%,Nei′s基因多样性指数（H）和Shannon信息指数（I）分别为0.1657和0.2448。在物种水平上,多态位点百分率（PPB）为79.31%,Nei′s基因多样性指数（H）和Shannon信息指数（I）分别为0.2947和0.4355。居群间的遗传分化系数(G_ST)达0.4349。结果表明：滇牡丹遗传多样性水平较高,居群间遗传分化较大。结合以前的研究结果,对滇牡丹的现状进行评估的结果显示,滇牡丹并不濒危。     

7种川产淫羊藿属植物遗传多样性及亲缘关系的ISSR分析

利用ISSR分子标记对7种12居群的川产淫羊藿属植物进行遗传多样性及亲缘关系分析。结果发现23个ISSR随机引物共扩增194条清晰条带,其中169条具多态性,平均多态性位点比率为87.11%,有效等位基因数Ne=1.534 2,Nei基因多样性指数H=0.314 4,Shannon多样性指数I=0.469 7,表明物种间遗传多样性丰富。7种川产淫羊藿属植物间的遗传相似系数为0.653 2～0.748 9,聚类分析和主成分分析直观的显示出12份供试材料间的亲缘关系,并将其分为两类。表明ISSR分子标记技术可以用于川产淫羊藿属植物的遗传多样性和亲缘关系分析。     

基于RAPD标记的丽江云杉遗传多样性及谱系地理

由于青藏高于独特的地理环境,第四纪冰期反复的退缩和扩展对目前该地区生物物种的地理分布及居群结构产生重要的影响。本文对这一地区特有分布物种丽江云杉(Picea likiangnsis)3个变种丽江云杉变种(var. likiangensis)、川西云杉变种(var. rubescens)和林芝云杉变种(var. linzhiensis)共11个种群228个个体进行RAPD分析,研究其遗传多样性及谱系地理。结果表明,该种具有较高的总体遗传多样性,多态位点达85.42%;然而在群体水平上却保持了相对低的多态性,种群间差异不显著,种群平均多态条带百分率为62.31%,Nei’s平均遗传多样性指数(H_E)为0.250,Shannon’s多样性指数(H_pop)从0.267到0.421 1。Nei’s基因多样性(G_ST=0.256)和AMOVE分析(Phi_st=0.236)表明,群体间有较高的遗传分化,群体间基因流有限(N_m=1.453 2),远远低于已报道的其它松科植物。UPGMA聚类分析表明3个形态上分离的变种没有单个聚成一枝,形成单系群。本研究认为,在第四纪冰期丽江云杉这一种在南部可能存在至少3个不同的避难所,北边和西边的居群应该是南方避难所里的居群经过不同的回迁路线而产生的,有可能是由于种内亚种间的反复杂交造成了目前的种群分布模式。     

野生罗汉果遗传多样性的ISSR分析

应用ISSR分子标记方法对采自广西和广东的7个罗汉果（Siraitia grosvenorii）野生居群共130个个体进行了遗传多样性分析。15个ISSR引物共扩增到了111个位点,其中91个是多态性位点,占82.0%。Nei′s基因多样性指数(H_e)为 0.248,Shannon 信息多样性指数（I） 为0.354。罗汉果不同居群的遗传多样性水平差异较大,居群多态位点百分率在 28.2%－55.6%之间,Nei′s基因多样性指数为0.080－0.209,Shannon 信息多样性指数为0.123－0.310。永福居群（YF）和金秀居群（JX）的遗传多样性水平较高,其周边居群的遗传多样性水平逐渐降低,居群间产生了较大的遗传分化（G_st = 0.569）。居群间的遗传距离与地理距离相关性不明显（r =0.369,P = 0.115）。UPGMA聚类图中,7个居群的个体按居群各自聚在一起。     

珍稀濒危植物长叶红砂种群遗传多样性的ISSR分析

采用ISSR分子标记技术,对濒危小灌木长叶红砂(Reaumuria trigyna) 集中分布的5个种群的遗传多样性水平和遗传结构进行了研究。14条引物共检测到114个位点,其中99个为多态位点,多态位点比率为86.84％,长叶红砂种群具有较高的遗传多样性。物种水平上Shannon多样性指数(I)为0.468 8,Nei基因多样性指数(H)为0.308 4;种群水平上,多态位点比率P为77.89%,I为0.410 6,H为0.260 9,基因分化系数Gst为0.106 9,揭示了长叶红砂种群遗传变异多存在于种群内,种群间的遗传分化较小,占10.69%。 基因流(Nm)为4.178 7＞1,说明种群间的基因交流,防止了由于遗传漂变导致的遗传分化。聚类分析表明长叶红砂种群遗传距离与地理距离之间无显著的相关性。研究结果说明遗传多样性水平与物种本身特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低。     

Genetic diversity of the endangered Chinese endemic plant Monimopetalum chinense revealed by amplified fragment length polymorphism (AFLP)

Monimopetalum chinense Rehd. is an endangered woody vine endemic to eastern China. Using amplified fragment length polymorphism (AFLP) markers, we examined levels of genetic variation within and among eleven populations located across the species’ distribution. Although modest levels of heterozygosity were detected, other measures of genetic diversity registered relatively high levels of variability, both at the species level (P = 91.0%, H_E = 0.232, I_S = 0.365) and at the population level (P = 53.0%, H_E = 0.155, I_S = 0.239). Populations also exhibited high levels of genetic differentiation (Nei’s genetic diversity analysis, G_ST = 0.330), corresponding to isolation-by-distance and hierarchical population structure. These results indicate that, despite low levels of gene flow, populations of M. chinense still harbor substantial amounts of genetic diversity. Management plans for the species should include measures that ensure genetic diversity remains high within and among extant populations.     

白沙蒿不同地理种群遗传分化的ISSR分析

运用ISSR技术,对白沙蒿的5个种群进行遗传分化的分析。12个引物在108个个体中共扩增出222个位点,其中,多态位点为218个,多态位点百分率为98.20%,Shannon多样性指数为0.315 4,具有高的多态性。种群间的遗传分化系数（G_st）为0.076 7,与AMOVA分析的结果一致（Ф_st为7.96%）,表明绝大多数的遗传变异存在于种群内部。各种群间遗传一致度高达98%以上,遗传距离很小,基因流达3.008 2,均表现出白沙蒿各种群存在着广泛的基因交流,有着很小的遗传分化。     

Population genetic diversity and divergence of the halobiotic herb Limonium sinense estimated by AFLP and ISSR, and implications for conservation

Limonium sinense is a halobiotic herb endemic to China that has been traditionally used for hundreds of years for its good restorative function. Genetic variation and population structure of this species were investigated by using amplified fragment length polymorphisms (AFLPs) and inter simple sequence repeats (ISSRs). A high level of genetic diversity was detected [AFLP: H _E = 0.284, percentage of polymorphic loci (PPL) = 92.68 %; ISSR: H _E = 0.257, PPL = 85.71 %] at the species level with POPGENE. Based on analysis of molecular variation (AMOVA), the among-population component accounted for 29.03 % (AFLP) and 28.81 % (ISSR) of the genetic variation, indicating that most of the genetic variation was between individuals within populations. The Shannon diversity index (I) was higher for AFLP (0.432) than for ISSR (0.395). Five main clusters were shown in the unweighted pair-group method with arithmetic mean (UPGMA) dendrogram created using TFPGA, consistent with the result of principal coordinate analysis using NTSYS. In situ conservation is advocated first. Keeping a stable environment for this halobiotic herb is necessary. For ex situ conservation, it is important to establish a germplasm bank. AFLP and ISSR markers were proved to be efficient tools in assessing the genetic variation among populations of L. sinense. The patterns of variation appeared to be consistent for these two marker systems, and they can be used for management of genetic structure, protection of the halobiotic plant, and conservation of germplasm.     

An AFLP-based survey of genetic diversity and relationships of major farmed cultivars and geographically isolated wild populations of <Emphasis Type="Italic">Saccharina japonica</Emphasis> (Phaeophyta) along the northwest coasts of the Pacific

The genetic diversity and relationships of six representative cultivars and six geographically isolated wild populations of Saccharina japonica along the northwest coasts of the Pacific Ocean were investigated using AFLP markers. A total of 547 bands were generated across all samples by ten primer combinations. At the cultivar or population level, the percentage of polymorphic loci (P), gene diversity (H), and Shannon’s information index (I) was highest in Dalian population (P 59.05%; H 0.2057; I 0.3062) and lowest in Lianjiang cultivar (P 9.87%; H 0.0331; I 0.0497). At the species level, P, H, and I were 85.01%, 0.1948, and 0.3096, respectively. Unique bands were detected in all the six wild populations, with Dalian being the most. In comparison, only Yanza cultivar possessed one unique band. The G _ST value was 0.6226 and the gene flow (N _m ) was 0.1515, indicating strong genetic differentiation among cultivars and populations. Two UMPGA dendrograms were constructed based on the Dice similarity coefficients among individuals and on genetic distances among cultivars and populations, which generally revealed three major clades corresponding to three countries. Analysis of molecular variance revealed that a larger proportion (60.21%) of the total genetic variation was attributable to differences among cultivars and populations. The Mantel test suggested that genetic differentiation was positively correlated with geographic distance (r = 0.7962, P = 0.011) in the six wild populations, agreeing with the isolation by distance model. On the whole, low to moderate genetic diversity within cultivars and populations (except Dalian population) and high genetic differentiation among cultivars and populations were detected.     

Genetic diversity of the endangered Chinese endemic herb Primulina tabacum (Gesneriaceae) revealed by amplified fragment length polymorphism (AFLP)

Primulina tabacum Hance, is a critically endangered perennial endemic to limestone area in South China. Genetic variability within and among four extant populations of this species was assessed using AFLP markers. We expected a low genetic diversity level of this narrowly distributed species, but our results revealed that a high level of genetic diversity remains, both at population level (55.5% of markers polymorphic, H _E = 0.220, I _S = 0.321), and at species level (P = 85.6% of markers polymorphic, H _E = 0.339, I _S = 0.495), probably resulting from its refugial history and/or breeding system. High levels of genetic differentiation among populations was apparent based on Nei’s genetic diversity analysis (G _st=0.350). The restricted gene flow between populations is a potential reason for the high genetic differentiation. The population genetic diversity of P. tabacum revealed here has clear implications for conservation and management. To maintain present levels of genetic diversity, in situ conservation of all populations is necessary.     

Conservation genetics of Heritiera littoralis (Sterculiaceae), a threatened mangrove in China,based on AFLP and ISSR markers

AFLP and ISSR markers were used to determine the genetic variations in eight mangrove and non-mangrove populations of Heritiera littoralis (Sterculiaceae), a threatened species in China. Our results showed a moderate to high level of genetic variation in this species (P = 63.69%, H_T = 0.20 for AFLP; P = 76.07%, H_T = 0.22 for ISSR), and a relatively high level of genetic differentiation among populations (G_ST = 0.24 for AFLP and 0.27 for ISSR). Life history traits, long-distance dispersal of floating seeds, and local environments may play important roles in shaping the genetic diversity and genetic structure of this species. Investigation of the plant’s reproductive capacity showed that the natural seed production of H. littoralis was low, as was the germination rate and the transformation rate from juvenile to adult. H. littoralis is seriously threatened and is in urgent need of conservation in China.     

Genetic diversity and structure of natural Pinus tabulaeformis populations in North China using amplified fragment length polymorphism (AFLP)

Chinese pine (Pinus tabulaeformis carr.), endemic to China, is a conifer species with extensive and fragmented distribution in North China. In this study, the genetic diversity and structure of 20 natural populations of this species were investigated using amplified fragment length polymorphism (AFLP) markers. A total of 445 fragments were revealed with 8 pairs of primers, 379 (85.17%) of which were polymorphic. A moderate level of genetic diversity was detected at the species level (Shannon's information index I = 0.356, Nei's gene diversity H_E = 0.271) and at the population level (I = 0.219, H_E = 0.206). Most of genetic variation was within populations while a considerable level of genetic differentiation was detected (G_ST = 0.352, Ф_ST = 0.304). The high differentiation could be attributed to the complex and fragmented habitats, and a limited gene flow among populations (N_m = 0.572). The Mantel test indicated that there was significant correlation (r = 0.455, P < 0.001) between Nei's genetic distance and geographical distance among all the populations. The results suggested that proper countermeasures should be taken to prevent the habitat further deterioration and maintain the genetic diversity of this species.     

硬核资源遗传多样性的AFLP分析

为了解云南省硬核[Scleropyrum wallichianum（Wight et Arn.）Arn.]的遗传多样性,采用AFLP标记分析了7个居群84份种质材料的遗传变异。结果表明,从64对引物组合中挑选出多态性较好的引物8对,共扩增出1 728条带,其中多态性条带1 388条,多态性百分率为80.14%。硬核在物种水平的多样性指数分别为Na=1.416,Ne=1.179,H=0.137,I=0.225,在居群水平上分别为H=0.111,I=0.175;在遗传相似性系数为0.52时,这些种质材料可分为3组,其中易武居群具有丰富的遗传变异,大部分的遗传变异存在于居群内,而在0.05置信区间内居群间遗传变异仅为11.5%;居群间的遗传距离和地理距离无显著相关性（r=0.0323,P=0.5820）。因此,硬核资源可采用就地和迁地保护策略,以增加其遗传多样性。     

Genetic diversity and relationship of endangered plant Magnolia officinalis (Magnoliaceae) assessed with ISSR polymorphisms

Twenty-eight populations of the rare medicinal plant Magnolia officinalis (Magnoliaceae) were sampled across its natural range, and inter-simple sequence repeats (ISSRs) markers were used to assess the genetic variation within and among populations. Twelve primer combinations produced a total of 137 unambiguous bands of which 114 (83.2%) were polymorphic. M. officinalis exhibited a relatively low genetic diversity at population level (the percentage of polymorphic loci PPB = 49.8%, Nei’s genetic diversity H = 0.194, Shannon’s information index I = 0.286). However, the genetic diversity at species level was relatively high (PPB = 83.2%; H = 0.342; I = 0.496). The coefficient of gene differentiation (G_ST, 42.8%) and the results of analysis of molecular variance (AMVOA) indicated that genetic differentiation occurred mainly within populations. The estimated gene flow (Nm) from G_ST was 0.669. It indicated that the fragmentation and isolation of populations might result from specific evolutionary history and anthropogenic activity. Genetic drift played a more important role than gene flow in the current population genetic structure of M. officinalis. Conservation strategies for this rare species are proposed based on the genetic data.